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Isoelectric focusing of alpha‐1 acid glycoprotein (orosomucoid) in immobilized pH‐gradients with 8 M urea: Detection of its desialylated variants using an alkaline phosphatase‐linked secondary antibody system
Author(s) -
Eap Chin B.,
Baumann Pierre
Publication year - 1988
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150091005
Subject(s) - orosomucoid , isoelectric focusing , chemistry , glycoprotein , urea , chromatography , isoelectric point , alkaline phosphatase , biochemistry , alpha (finance) , enzyme , medicine , construct validity , nursing , patient satisfaction
A method allowing a clear separation of the different variants of desialylated alpha 1‐acid glycoprotein (orosomucoid) has been developed using isoelectric focusing in immobilized pH gradients, supplemented with 8 M urea and 2 % v/v 2‐mercaptoethanol. Immunoblotting with two antibody‐steps afforded high sensitivity and permitted the detection of about 700 pg of alpha 1‐acid glycoprotein in a 20 μL plasma sample diluted 1:28 672. A one year old bloodstain, kept at room temperature, could easily be phenotyped.